Genome-scale DNA methylation mapping of clinical samples at single-nucleotide resolution

被引：224

作者：

Gu, Hongcang ^{[1
]}

Bock, Christoph ^{[1
,2
,3
,4
]}

Mikkelsen, Tarjei S. ^{[1
]}

Jaeger, Natalie ^{[1
,2
,3
]}

Smith, Zachary D. ^{[1
,2
,3
]}

Tomazou, Eleni ^{[1
,2
,3
]}

Gnirke, Andreas ^{[1
]}

Lander, Eric S. ^{[1
,5
,6
]}

Meissner, Alexander ^{[1
,2
,3
]}

机构：

[1] Broad Inst, Cambridge, MA USA

[2] Harvard Univ, Dept Stem Cell & Regenerat Biol, Cambridge, MA 02138 USA

[3] Harvard Stem Cell Inst, Cambridge, MA USA

[4] Max Planck Inst Informat, Saarbrucken, Germany

[5] MIT, Dept Biol, Cambridge, MA USA

[6] Harvard Univ, Sch Med, Dept Syst Biol, Boston, MA USA

来源：

NATURE METHODS | 2010年 / 7卷 / 02期

基金：

美国国家卫生研究院;

关键词：

COLORECTAL-CANCER; MAPS; EPIGENOME; COLON;

D O I：

10.1038/nmeth.1414

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Bisulfite sequencing measures absolute levels of DNA methylation at single-nucleotide resolution, providing a robust platform for molecular diagnostics. We optimized bisulfite sequencing for genome-scale analysis of clinical samples: here we outline how restriction digestion targets bisulfite sequencing to hotspots of epigenetic regulation and describe a statistical method for assessing significance of altered DNA methylation patterns. Thirty nanograms of DNA was sufficient for genome-scale analysis and our protocol worked well on formalin-fixed, paraffin-embedded samples.

引用

页码：133 / U69

页数：6

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